AbstractAspergillus niger NCIM 1207 produced significantly high levels of β-glucosidase and β-xylosidase activities in submerged fermentation. Cellulose induced only β-glucosidase, while xylan induced both β-glucosidase and β-xylosidase activities. Both the enzymes of this strain were found to undergo catabolite repression in the presence of high concentrations of glucose and glycerol. The sudden drop in pH of the fermentation medium below 3.5 caused the inactivation of enzymes when the fungus was grown in glycerol-containing media at lower temperatures. The growth of the organism at 36 oC led to an increase in pH of the fermentation above 6.0 that affected β-xylosidase activity significantly. Highest levels of β-glucosidase ((19 IU mL-1 or 633 IU g-1 of substrate) and β-xylosidase (18.7 IU/mL-1 or 620 IU g-1 of substrate) activities were detected when A. niger was grown at 30 oC for first five days followed by further incubation at 36 oC. Such a process of growing the organism at lower temperatures (growth phase) followed by producing the enzymes at higher temperatures (production phase) in case of fungal systems has not been reported so far. The zymogram staining of the β-glucosidase demonstrated that A. niger produced only single species of β-glucosidase. We feel that A. niger NCIM 1207 is a potential candidate to produce both β-glucosidase and β-xylosidase in high amounts that can be used to supplement commercial cellulase preparation.